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Published Online: 12 March 2021

Characterization of lsa(D), a Novel Gene Responsible for Resistance to Lincosamides, Streptogramins A, and Pleuromutilins in Fish Pathogenic Lactococcus garvieae Serotype II

Publication: Microbial Drug Resistance
Volume 27, Issue Number 3


Aims: Fish pathogenic Lactococcus garvieae serotype II has been isolated from cultured fish species in Japan. This study aimed to investigate the molecular mechanisms of lincomycin (LCM)-resistant L. garvieae serotype II and assess the molecular basis for lincosamides–streptogramins A–pleuromutilins (LSAP)-resistant phenotype.
Results: We identified a novel lsa(D)-encoded 497-aa ATP-binding cassette F (ABC-F) protein in the LSAP-resistant strains. Amino acid identities of 41.25–54.73% were obtained between the deduced amino acids from Lsa(D) and other Lsa-type ABC-F proteins. Furthermore, comparative analysis revealed that the allele of lsa(D) with single point mutation at 233 aa position (TGG → TAG; tryptophan→premature termination codon [PTC]) in LSAP-sensitive strains. The minimum inhibitory concentrations of antimicrobials against the lsa(D) complementary strain and lsa(D)-disrupted mutant confirmed that lsa(D) conferred the LSAP-resistant phenotype. The reverse transcription–polymerase chain reaction could not detect the noncoding region of lsa(D) allelic variant in the LSAP-sensitive strains. Additionally, the PTC (TAG) in LCM-sensitive strains was replaced by TGG, CAG, or TAT in the laboratory-induced revertant mutants.
Conclusions: The novel lsa(D) conferred the LSAP-resistant phenotype in clinically LCM-resistant L. garvieae serotype II strains. However, the allele of lsa(D) gene containing the PTC was found in L. garvieae serotype II, resulting in the LSAP-susceptible phenotype.

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Published In

cover image Microbial Drug Resistance
Microbial Drug Resistance
Volume 27Issue Number 3March 2021
Pages: 301 - 310
PubMed: 32706619


Published online: 12 March 2021
Published in print: March 2021
Published ahead of print: 20 July 2020


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Faculty of Agriculture, University of Miyazaki, Miyazaki, Japan.
Terutoyo Yoshida
Faculty of Agriculture, University of Miyazaki, Miyazaki, Japan.
Atushi Fujiwara
Research Center for Bioinformatics and Biosciences, National Research Institute of Fisheries Science, Fisheries Research and Education Agency, Yokohama, Japan.
Issei Nishiki
Research Center for Bioinformatics and Biosciences, National Research Institute of Fisheries Science, Fisheries Research and Education Agency, Yokohama, Japan.


Address correspondence to: Yin-Ze Shi, MS, Faculty of Agriculture, University of Miyazaki, Miyazaki 889–2192, Japan [email protected]

Disclosure Statement

No competing financial interests exist.

Funding Information

This study was supported by a Grant-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology, Japan (18K05825).

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